Purification And Functions Of Bio-Active Materials From Protaetia Brevitarsis (Coleoptera) Larva And Phellinus Baumii

Phellinus fungus is a member of Hymenochaetaceae,which have been used as a traditional medicine in oriental countries for the treatment of tumors,gastroenteric disorders,inflammation and lymphatic diseases.For its anti-tumor activity,Phellinus fungus also have lots of advantages as an anti-cancer candidate.Part one of this paper shows that finding active matial from Phellinus baumii.We has found a novel lectin,a novel anti-fungi molecular and a water-soluble polysaccharide.In this experiment,A novel lectin from the liquid cultured mycelia of medicinal fungus Phellinus baumii was purified by ion chromatography and HPLC.The relative molecular mass of it was about 24 kDa tested by HPLC and SDS-PAGE.This implied it was composed of one subunit.GAL could agglutinated red blood cells of mouse and people, while its hemagglutinating activity for different resources of red blood cells was different.It can't be inhibited by D-fructose,D-galactose,dextrose,cane sugars,lactose.GAL was thermo-stabIe,and metal cations such as Ca²⁺,Mg²⁺ and Zn²⁺ had no effects on it.With the same method,we got a new molecular,the mass of it was about 20 kDa.It owns anti-fungi activity.The Phellinus mushroom has been known to exhibit various potent biological activities.Little is known about the polysaccharide extracted from Phellinus baumii. Rencently,a water-soluble polysaccharide was isolated from the mycelia of Phellinus baumii by hot water extraction,free proteins were removed by Sevage method.The process of purifying this crude polysaccharide was achieved by anion-exchange and gel-permeation chromatography.Acidic fractions of the polysaccharide were separated from crude polysaccharides by DEAE-Sephadex A-50 anion exchange chromatography at 1.1 M NaCI.The molecular weight of this polyaccharide(PBG) after Sephadex G-75 was about 17,000D.By testing the activity,PBG was found to increase the proliferation of RAW264.7 cells markedly.At the same time,PBG could inhibit the proliferation of HepG2 cells and HCT cells largely.Part one of this paper shows that a novel water-soluble polysaccharide has been found from Protaetia brevitarsis larva(grub).With the same method,the crude polysaccharides were extracted from one medical insect,Protaetia brevitarsis larva(grub),by homogenization,ethanol and Sevag method. The proliferation effects of polysaccharides of grub on immunological ceils were estimated by MTT assay,which indicated that the crude polysaccharide of grub could significantly improve the proliferation of splenocytes and macrophages of mice.The amount of total carbohydrates of grub polysaccharides was measured by phenol-sulfuric acid method.The crude polysaccharides was isolated and purified by using DEAE-SephadexA-25,LPLC-Sephadex G-75 column.Two kinds of polysaccharides were associated with the activity to improve mice splenocytes proliferation in vitro,the molecular weight of which were 42kDa,26 kDa separately.